白藜芦醇基于Notch1/Hes1信号通路对丙戊酸诱导孤独症模型小鼠重复呆板样和记忆衰退症状的影响

The Effect of Resveratrol on Valproic Acid-induced Repeated Dullness and Memory Decline Symptoms in Autism Rats Based on Notch1/Hes1 Pathway

目的探讨白藜芦醇基于Notch1/Hes1信号通路对丙戊酸(VPA)诱导孤独症小鼠的重复呆板样和记忆衰退症状的影响。方法健康成年C57BL/6J小鼠,雄性10只,雌性20只,合笼饲养。受孕后雌鼠随机分为两组:模型组(n=10),于E13 d腹腔注射VPA 600 mg·kg^(-1)对照组(n=6),于E13 d腹腔注射等量0.9%氯化钠溶液(NS)。模型组雌鼠所产雄性子鼠随机分为3组:VPA+白藜芦醇组(n=7):皮下注射白藜芦醇3.6 mg·kg^(-1),连续给药13 d;VPA+NS组(n=8):皮下注射等量0.9%NS;VPA组(n=6),未经任何处理。对照组雌鼠所产雄性子鼠设为NS组(n=8)。利用VPA构建孤独症模型,三箱实验鉴定造模成功,采用旷场实验和Morris水迷宫实验检测模型小鼠的重复呆板样行为和记忆能力;Western blot法检测小脑和海马Notch1/Hes1信号通路蛋白Notch1、Jagged1、NICD及Hes1的表达水平。结果本研究成功构建孤独症模型。与NS组比较,VPA组模型小鼠社会交互能力下降(P<0.01),重复呆板样行为增加(P<0.05),穿越平台次数及在靶象限的时间减少(P<0.05),小脑及海马脑区的Notch1、Jagged1、NICD和Hes1蛋白表达均升高(均P<0.05);白藜芦醇干预后,与VPA+NS组比较,VPA+白藜芦醇组模型小鼠重复呆板样行为减少(P<0.05),穿越平台次数及在靶象限的时间均增加(均P<0.05),小脑及海马脑区的Notch1、Jagged1、NICD和Hes1蛋白表达均降低(均P<0.05)。结论白藜芦醇可改善VPA诱导孤独症模型小鼠的重复呆板样与记忆衰退症状,降低Notch1/Hes1信号通路相关蛋白的过度活化可能是其作用机制之一。

Aim To investigate the effect of resveratrol on the repeated dullness and memory decline in valproic acid(VPA)-induced autism rats based on Notch1/Hes1 pathway.Methods Healthy adult C57BL/6J(C57)mice,including 10 males and 20 females,were reared in cages.After conception,female mice were randomly divided into two groups.The model group(n=10)was intraperitoneally injected with VPA 600 mg·kg^(-1)on E13 d.The control group(n=6)was intraperitoneally injected with 0.9%sodium chloride solution(NS)on E13d.Male offspring of the model group were randomly divided into 3 groups:a VPA+resveratrol group(n=7),3.6 mg·kg^(-1)resveratrol was subcutaneously administered for 13 days;a VPA+NS group(n=8),subcutaneous injection of 0.9%NS;a VPA group(n=6),without any treatment.Male offspring of female mice in the control group were set as a NS group(n=8).VPA was used to construct the autism model,and the open field test and Morris water maze test were used to test the repeated mechanical behavior and memory ability of the model mice after the three box test.Western blot was used to detect the expression levels of Notch1,Jagged1,NICD and Hes1 in cerebellum and hippocampus.Results The rat model of autism was established successfully.Compared with the NS group,the social interaction ability(P<0.01)reduced,repetitive/stereotypic-like movements(P<0.05)increased,the number of crossing platform and the time spent in the platform(P<0.05)decreased,the protein level of Notch1,Jagged1,NICD,and Hes1 in the cerebellar and hippocampal brain regions(P<0.05)increased in the VPA group.However,compared with the VPA+NS group,repetitive/stereotypic-like movements(P<0.05)reduced,the number of crossing platform and the time spent in the platform(P<0.05)increased,the protein level of Notch1,Jagged1,NICD,and Hes1 in cerebellar brain area and hippocampal brain area(P<0.05)reduced in the resveratrol group.Conclusion The benefits of resveratrol in repeated dullness and memory decline of autism model may be mediated by attenuating the Notch1/Hes1 pathway activity.